There is huge autofluorescence in red spectral region. We have tried autofluorescence-quenching using application of ammonium chloride, or ammonium acetate with copper (II) sulfate. But this solutions are useful mainly against lysozime autofluorescence which manifests itself in distinct spectral range.
Have you considered these old discussions in Researchgate?
https://www.researchgate.net/post/Do_you_have_any_suggestion_to_eliminate_red_blood_cells_autofluorescence_present_in_PFA-fixed_sections
https://www.researchgate.net/post/Does_anyone_have_a_good_protocol_to_reduce_remove_autofluorescence_in_tissues
Good Luck.
a robust method for dual ISH /IHC to generate concordant results is discussed here. check for your specific targets in search on the website below...
https://acdbio.com/science/applications/research-areas/inflammation
Thanks a lot! We have tried using of several quenching solutions (ammonium chloride, ammonium acetate + CuSO4, glycine, Sudan black, Pontamine sky blue, ..) but as the only way we see PBS perfusion previous fixation (Zamboni´s fixative containing formaldehyde instead of PFA + picric acid) and subsequent sample clearing by xylene. I'll let you know about result as soon as we will get it :-) The procedure described I have found in article
https://www.ncbi.nlm.nih.gov/pubmed/25549834
Regards
Lucie
https://www.ncbi.nlm.nih.gov/pubmed/25549834
- Badges
- Science topic
- Similar topics
- Physical Sciences
- Physical Phenomena
- Luminescence
- Fluorescence